Premature Stop Codon at Residue 101 within HIV-1 Rev Does Not Influence Viral Replication of Clade BC but Severely Reduces Viral Fitness of Clade B

Abstract

HIV-1 Rev is an accessory protein that plays a key role in nuclear exportation, stabilization, and translation of the viral mRNAs. Rev of HIV-1 clade BC often shows a truncation of 16 AAs due to a premature stop codon at residue 101. This stop codon presents the highest frequency in clade BC and the lowest frequency in clade B. In order to discover the potential biological effect of this truncation on Rev activity and virus replication of clade BC, we constructed Rev expression vectors of clade BC with or without 16 AAs within C-terminal separately, and replaced the stop codon by Q in a CRF07_BC infectious clone. We found that 16 AAs truncation had no effect on expression and activity of Rev in clade BC. Also, the mutation from the stop codon to Q had no effect on virus replication of clade BC. Next, to investigate the effect of this truncation on Rev activity and replication capacity of clade B, Rev expression vectors of clade B carrying or lacking 16 AAs in C-terminal were constructed respectively, and residue Q at position 101 within Rev was substituted by the stop codon in a clade B infectious clone. It was found that 16 AAs truncation significantly down-regulated Rev expression and impaired clade B Rev activity. Furthermore, a Q-to-stop codon substitution within Rev significantly reduced viral replication fitness of clade B. These results indicate that the premature stop codon at residue 101 within Rev exerts diverse impact on viral replication among different HIV-1 clades.

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Acknowledgements

We appreciate the generous gifts of AT1 and AT2 plasmids from Dr. Carrie Dykes (The University of Rochester School of Medicine and Dentistry), as well as BH10-Rev expression plasmid and pCMV-Gag-Pol-RRE expression plasmid of clade B from Dr. Min Wei (Nankai University). Clade BC Gag-Pol-RRE expression vector was received from Dr. Jingwan Han (Beijing Institute of Microbiology and Epidemiology). CMV-EGFP plasmid was gifted from Dr. Lei Yu (Shenzhen Second People’s Hospital). We would like to thank Hong Peng (The Division of Research of Virology and Immunology, National Center for AIDS/STD Control and Prevention) for the technical supports for this work. This study was partially funded by the National Natural Science Foundation of China (Grants 81872680 and 31600734), the Yong Scientific Research Foundation of NCAIDS/STD (Grant 2018AFQN002) and the SKID outstanding youth grant (2019SKLID402). Funding has no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript.

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ZW and YS conceived and designed the experiments. ZW and XJ performed the experiments. ZW, XJ and KH analyzed the data. DL, YH and YS contributed reagents/materials/analysis tools. ZW, KH and LM wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Zheng Wang or Yiming Shao.

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The authors declare that they have no conflict of interest.

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Informed consent have been obtained from all participants and the studies have been approved by the Institutional Review Board of the National Center for AIDS/STD control and Prevention, China CDC.

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Wang, Z., Ji, X., Hao, Y. et al. Premature Stop Codon at Residue 101 within HIV-1 Rev Does Not Influence Viral Replication of Clade BC but Severely Reduces Viral Fitness of Clade B. Virol. Sin. 35, 181–190 (2020). https://doi.org/10.1007/s12250-019-00179-0

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Keywords

  • HIV-1
  • CRF_BC
  • Rev
  • Premature stop codon
  • Replication